Development of a gene therapy for congenital phagocyte disorder using both genome editing and iPS cell technology

• Project/Area Number: 15H04874
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Pediatrics
• Research Institution: Shinshu University
• Principal Investigator: Koike Kenichi 信州大学, 医学部, 特任教授 (40143979)
• Co-Investigator(Kenkyū-buntansha): 中沢 洋三 信州大学, 医学部, 教授 (60397312) ; 重村 倫成 信州大学, 学術研究院医学系(医学部附属病院), 講師 (70623916)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥17,030,000 (Direct Cost: ¥13,100,000、Indirect Cost: ¥3,930,000); Fiscal Year 2017: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2016: ¥7,540,000 (Direct Cost: ¥5,800,000、Indirect Cost: ¥1,740,000); Fiscal Year 2015: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
• Keywords: iPS細胞 / ゲノム修飾 / 先天性好中球減少症 / 遺伝子治療 / 再生医学 / 白血病発症 / 先天性食細胞異常症

Outline of Final Research Achievements

In parallel with genetic editing experiments of iPS cells established from a patient with ELANE-mutated severe congenital neutropenia, we performed genetic editing experiments on iPS cells derived from a patient with juvenile myelomonocytic leukemia (JMML). Both iPS cells with PTPN11 gene mutation and those with wild type of PTPN11gene were established from a JMML patient. Mutant iPS cells possessed significantly higher ability to generate CD34 positive cells. In addition, whole-exome sequencing and Sanger sequencing revealed that JMML iPSCs with PTPN11 mutation possessed OSBP2 mutation, while JMML iPSCs with no PTPN11 mutation did not carry OSBP2 mutation. By homologous recombination using zinc finger nuclease, we corrected PTPN11 mutation in iPSCs with both PTPN11 mutation and OSBP2 mutation, and transduced PTPN11 mutation into JMML iPSCs with wild type of the two genes. The results suggest that PTPN11 mutation itself contributes to leukemogenesis of this patient.

Research Products

• [Journal Article] Panobinostat inhibits the proliferation of CD34+CD38－cells under stimulation of hematopoietic growth factors on AGM-S3 cells in juvenile myelomonocytic leukemia. (2018)
  • Author(s): Kurata, T, Matsuda K, Hirabayashi K, Shigemura T, Sakashita K, Nakahata T, Koike K.
  • Journal Title: Pediatr Blood Cancer Volume: 印刷中 Issue: 11
  • DOI: 10.1002/pbc.27261
  • Peer Reviewed / Open Access
• [Journal Article] Increasing Risk of Disturbed Root Development in Permanent Teeth in Childhood Cancer Survivors Undergoing Cancer Treatment at Older Age. (2017)
  • Author(s): Tanaka M, Kamata T, Yanagisawa R, Morita D, Saito S, Sakashita K, Shiohara M, Kurita H, Koike K, Nakazawa Y.
  • Journal Title: J Pediatr Hematol Oncol. Volume: 39 Issue: 3 Pages: e150-e154
  • DOI: 10.1097/mph.0000000000000788
  • Peer Reviewed
• [Journal Article] Dramatic Reduction in Tumor Size During 5 Months of Pazopanib Therapy in Combination With Ifosfamide, Carboplatin, and Etoposide in an Early Infant With Progressive Soft Tissue Sarcoma. (2017)
  • Author(s): Watanabe T, Kurata T, Sano K, Suzuki S, Kaneko T, Motobayashi M, Shigemura T, Sumi T, Koike K, Nakazawa Y
  • Journal Title: J Pediatr Hematol Oncol Volume: 39 Issue: 2 Pages: 154-6
  • DOI: 10.1097/mph.0000000000000628
  • Peer Reviewed
• [Journal Article] IFR4/MUM1-positive lymphoma in Waldeyer ring with co-expression of CD5 and CD10. (2017)
  • Author(s): Chen L, Al-Kzayer LF, Liu T, Kobayashi N, Nakazawa Y, Koike K.
  • Journal Title: Pediatr Blood Cancer Volume: 64 Issue: 2 Pages: 311-314
  • DOI: 10.1002/pbc.26236
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Aberrant methylation of protocadherin 17 and its prognostic value in pediatric acute lymphoblastic leukemia (2017)
  • Author(s): Uyen TN, Sakashita K, Al-Kzayer LF, Nakazawa Y, Kurata T, Koike K.
  • Journal Title: Pediatric Blood & Cancer Volume: in press Issue: 3 Pages: 200-5
  • DOI: 10.1002/pbc.26259
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Clinical utility of next-generation sequencing for inherited bone marrow failure syndromes. (2017)
  • Author(s): Muramatsu H, Okuno Y, Yoshida K, Shiraishi Y, Doisaki S, Narita A, Sakaguchi H, Kawashima N, Wang X, Xu Y, Chiba K, Tanaka H, Hama A, Sanada M, Takahashi Y, Kanno H, Yamaguchi H, Ohga S, Manabe A, Harigae H, Kunishima S, Ishii E, Kobayashi M, Koike K, Watanabe K, Ito E, Takata M, Yabe M, Ogawa S, Miyano S, Kojima S.
  • Journal Title: Genet Med. Volume: - Issue: 7 Pages: 796-802
  • DOI: 10.1038/gim.2016.197
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Increasing Risk of Disturbed Root Development in Permanent Teeth in Childhood Cancer Survivors Undergoing Cancer Treatment at Older Age. (2017)
  • Author(s): Tanaka M, Kamata T, Yanagisawa R, Morita D, Saito S, Sakashita K, Shiohara M, Kurita H, Koike K, Nakazawa Y.
  • Journal Title: J Pediatr Hematol Oncol. Volume: 39
  • Peer Reviewed / Open Access
• [Journal Article] Pazopanib Therapy in Combination With Ifosfamide, Carboplatin, and Etoposide in an Early Infant With Progressive Soft Tissue Sarcoma. (2017)
  • Author(s): Watanabe T, Kurata T, Sano K, Suzuki S, Kaneko T, Motobayashi M, Shigemura T, Sumi T, Koike K, Nakazawa Y.
  • Journal Title: J Pediatr Hematol Oncol Volume: 39 Pages: 154-156
  • Peer Reviewed / Open Access
• [Journal Article] Viral load and ganciclovir (GCV) concentration in cerebrospinal fluid of patients successfully treated with GCV or valGCV for human herpesvirus 6 encephalitis/myelitis following umbilical cord blood transplantation. (2016)
  • Author(s): D. Morita, K. Hirabayashi, Y. Katsuyama, H. Morokawa, M. Motobayashi, T. Kurata, T. Shigemura, M. Tanaka, Y. Inaba, K. Koike, Y. Nakazawa.
  • Journal Title: Transpl Infect Dis Volume: 18 Issue: 5 Pages: 773-776
  • DOI: 10.1111/tid.12579
  • Peer Reviewed / Open Access
• [Journal Article] Diagnosis and Treatment of Juvenile Myelomonocytic Leukemia (2016)
  • Author(s): Sakashita K, Matsuda K, Koike K
  • Journal Title: Pediatr Int Volume: 58 Issue: 8 Pages: 681-90
  • DOI: 10.1111/ped.13068
  • Peer Reviewed / Open Access
• [Journal Article] Loss of Mismatched HLA on the Leukemic Blasts of Patients With Relapsed Lymphoid Malignancies Following Bone Marrow Transplantation From Related Donors With HLA Class II Mismatches in the Graft Versus Host Direction. (2016)
  • Author(s): Hirabayashi K, Kurata T, Horiuchi K, Saito S, Shigemura T, Tanaka M, Yanagisawa R, Matsuda K, Sakashita K, Koike K, Nakazawa Y
  • Journal Title: Pediatr Blood Cancer Volume: 63 Issue: 4 Pages: 709-11
  • DOI: 10.1002/pbc.25819
  • Peer Reviewed / Open Access
• [Journal Article] Anti-proliferative effects of T cells expressing a ligand-based chimeric antigen receptor against CD116 on CD34(+) cells of juvenile myelomonocytic leukemia (2016)
  • Author(s): Nakazawa Y, Matsuda K, Kurata T, Sueki A, Tanaka M, Sakashita K, Imai C, Wilson MH, Koike K
  • Journal Title: J Hematol Oncol Volume: 9 Issue: 1 Pages: 27-27
  • DOI: 10.1186/s13045-016-0256-3
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Intrathecal donor lymphocyte infusion for isolated leukemia relapse in the central nervous system following allogeneic stem cell transplantation: a case report and literature review. (2016)
  • Author(s): Yanagisawa R, Nakazawa Y, Sakashita K, Saito S, Tanaka M, Shiohara M, Shimodaira S, Koike K.
  • Journal Title: Int J Hematol Volume: 103 Issue: 1 Pages: 107-11
  • DOI: 10.1007/s12185-015-1902-1
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Analysis of KRAS and NRAS gene mutations in Arab Asian children with acute leukemia: High frequency of RAS mutations in acute lymphoblastic leukemia. (2015)
  • Author(s): Al-Kzayer LF, Sakashita K, Al-Jadiry MF, Al-Hadad SA, Ghali HH, Uyen LT, Liu T, Matsuda K, Abdulkadhim JM, Al-Shujairi TA, Matti ZI, Sughayer MA, Rihani R, Madanat FF, Inoshita T, Kamata M, Koike K.
  • Journal Title: Pediatr Blood Cancer Volume: 62 Issue: 12 Pages: 2157-61
  • DOI: 10.1002/pbc.25683
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Subcutaneous abscess due to the basidiomycete Phellinus mori in a patient with chronic granulomatous disease. (2015)
  • Author(s): Shigemura T, Nakazawa Y, Amano Y, Sudo A, Watanabe M, Kobayashi M, Kobayashi N, Koike K, Agematsu K, Nishimura K.
  • Journal Title: Infection Volume: 43 Issue: 3 Pages: 371-5
  • DOI: 10.1007/s15010-015-0724-7
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Dramatic improvement in the multifocal positron emission tomography findings of a young adult with chronic granulomatous disease following allogeneic hematopoietic stem cell transplantation. (2015)
  • Author(s): Shigemura T, Nakazawa Y, Hirabayashi K, Kobayashi N, Sakashita K, Agematsu K, Koike K.
  • Journal Title: J Clin Immunol Volume: 35 Issue: 1 Pages: 84-6
  • DOI: 10.1007/s10875-014-0113-5
• [Presentation] 臍帯血移植後の輸血関連サイトメガロウイルス血症の検討 (2018)
  • Author(s): 重村倫成、小森一寿、盛田大介、平林耕一、倉田敬、齋藤章治、田中美幸、柳沢龍、坂下一夫、中沢洋三
  • Organizer: 第40回日本造血細胞移植学会総会
• [Presentation] 重症先天性好中球減少症責任遺伝子ELANE変異を認めた無症候性母親の解析 (2018)
  • Author(s): 重村倫成、小林法元、上松一永、中沢洋三、小原収
  • Organizer: 第1回日本免疫不全・自己炎症学会
• [Presentation] A novel heterozygous C243Y A20/TNFAIP3 gene mutation is responsible for chronic inflammation in autosomal dominant Behcet’s disease (2017)
  • Author(s): Shigemura T, Kobayashi N, Kobayashi K, Agematsu K
  • Organizer: 第60回小児リウマチ学会総会・学術集会
• [Presentation] Development of T cells redirected to GM-CSF receptor with a ligand-based CAR for myeloid neoplasms (2016)
  • Author(s): Nakano T, Morita D, Matsuda K, Ohnota H, Hongo K, Koike K, Nakazawa Y
  • Organizer: 日本血液学会
  • Place of Presentation: パシフィコ横浜（神奈川県）
  • Year and Date: 2016-10-13
• [Presentation] JMMLのCD34陽性細胞増殖に対するPanobistatとazacitidineのin vitro効果 (2015)
  • Author(s): 倉田 敬、小池 健一、他
  • Organizer: 第57回日本小児血液・がん学会学術集会
  • Place of Presentation: 甲府富士屋ホテル（山梨県甲府市）
  • Year and Date: 2015-11-27
• [Presentation] 体細胞ヘテロ接合性CBL変異が長期に残存したJMML (2015)
  • Author(s): 平林真介、小池 健一、他
  • Organizer: 第57回日本小児血液・がん学会学術集会
  • Place of Presentation: 甲府富士屋ホテル（山梨県甲府市）
  • Year and Date: 2015-11-27
• [Presentation] 臍帯血移植を行ったmucolipidosis II alpha/beta型の臨床経過 (2015)
  • Author(s): 柴崎拓美、小池 健一、他
  • Organizer: 第57回日本小児血液・がん学会学術集会
  • Place of Presentation: 甲府富士屋ホテル（山梨県甲府市）
  • Year and Date: 2015-11-27